Genotoxicity evaluation of drinking water sources in human peripheral blood lymphocytes using the comet assay

The potential harm of organic pollutants in drinking water to human health is widely focused on in the wodd; more and more pollutants with genotoxic substances are released into the aquatic environment. Water source samples were collected from 7 different localities of Nanjing City. The potential genotoxicity of organic extracts from drinking water sources were investigated by means of the comet assay in human peripheral lymphocytes. The results showed that all the organic extracts from all the water source samples could induce DNA damages of human peripheral blood lymphocytes at different levels. A significant difference （P 〈 0.01） was observed when compared with the solvent control, The DNA damage increased with the increase of the dosage of the original water source. Significant differences of DNA damage were observed in different drinking water sources, as shown by the multiple comparisons analysis at the dosage of 100x; the degree of DNA damage treated by Hushu waterworks （at town level） was the most serious, the arbitrary units （AU） was 141.62±6.96, however, that of Shangyuanmen waterworks （at city level） was only 109.64±2.97. The analysis also revealed that the genotoxicity of town＇s water sources was higher than that of the city. The results demonstrated that the comet assay can be successfully applied to the genotoxicity monitoring programs of drinking water sources.

Metallothionein 1 Isoform Gene Expression Induced by Cadmium in Human Peripheral Blood Lymphocytes

Objective To study the gene expression of metallothionein 1 （MT-1） isoforms in human peripheral blood lymphocytes （HPBLs）. Methods The expression of mRNA representing the seven active MT-I genes was determined in HPBLs by quantitative RT-PCR before and after exposure to cadmium. Results Basal expressions of MT-IX, and MT-1A in HPBLs were similar to expression of housekeeping gene. In contrast, the basal gene expressions of MT- 1 H, IF, 1E, and 1G were a little transcripts in human HPBLs. No signal was detected for MT-1B. There was a sex difference （P〈0.05）. in basal gene expression of MT-1E. The levels of gene expression of MT-1A, 1E, IF, 1G, 1H, and 1X increased, but the level of MT-1B did not increase after exposure to cadmium. Conclusions Gene expressions of MT-1 G, MT-1 H, MT-1 F, and MT-1X in HPBLs can be used as a potential biomarker of cadmium exposure.

X-ray-induced Expression Changes of TNFSF4 Gene in Human Peripheral Blood

This study examined ionizing radiation-induced tumor necrosis factor （ligand） superfamily, member 4 （TNFSF4） mRNA expression changes in human peripheral blood cells and their distribution in a normal population. The results showed that expression level of TNFSF4 mRNA exhibited a dose- dependent response after different irradiation doses, but that was independent of incubation time post-irradiation. Moreover, it was not affected by age and gender in 51 healthy donors. Our studies indicate that TNFSF4 can be considered as a candidate gene to develop a new biodosimeter.

Polysaccharide from Astragalus membranaceus promotes the activation of human peripheral blood and mouse spleen dendritic cells

Astragalus membranaceus(A.membranaceus)is a widely used traditional herb in China and Korea.A.membranaceus polysaccharides(AMP),which make up a major part of the root extract,have been shown to modulate immune modulations,especially activation of bone marrow-derived dendritic cells(BMDCs)and T cells.However,the immune stimulatory effect of AMP in the mouse in vivo and human peripheral blood DCs(PBDCs)has not been well investigated.In this study,we found that intravenous(i.v.)injection of AMP in C57 BL/6 mice induced remarkable elevations in co-stimulatory and MHC class I and II molecule levels in the splenic DCs and its subsets.The stimulatory effect of DCs by AMP was elevated 6 h after treatment,which rapidly decreased 18 h after injection.Furthermore,AMP promoted intracellular production of pro-inflammatory cytokines in spleen DC subsets,which contributed elevation of serum cytokine levels.Finally,the AMP promoted PBDC activation.Thus,these results demonstrate that AMP can be used as an immune stimulatory molecules in human and mouse.

Investigation of the cytotoxicity,antioxidative and immune-modulatory effects of Ligusticum porteri(Osha) root extract on human peripheral blood lymphocytes

OBJECTIVE： Ligusticum ported is a traditional Native American herb. The roots of L. ported are traditionally used in the treatment of many diseases, however, its cytotoxicity, antioxidative and immune- modulatory effects need to be investigated. In this study, we evaluated the effects of the root extract at different doses on human peripheral blood lymphocytes （PBLs）. METHODS： The lymphocytes were incubated with different concentrations of the root extracts （0, 50, 100, 200, and 400 μg/mL） and harvested every 6 h for 2 d （P〈0.05）. The protective effect of the herb against oxidative damage was determined by inducing oxidative stress with the administration of 50 μmol/L of hydrogen peroxide （H202）. RESULTS： Treatments with L. ported at 200 and 400 pg/mL increased the viability of PBLs. The deleterious effect of H2O2 was ameliorated by 400μg/mL L. ported treatment. Addition of 400 μg/mL L. ported reduced lipid peroxidation in stressed PBLs by 94% （P〈0.05）. Treatment with 400 μg/mL of L. ported resulted in a 26.4% increase of reduced glutathione levels. Activities of superoxide dismutase and catalase increased by 17.5% and 55.2% respectively, when stressed PBLs were treated with 400 μg/mL L. ported for 2 d （P〈0.05）. Treatment with 400 μg/mL L. ported increased interferon-γand interleukin-2 expressions in H2O2-challenged PBLs （P〈0.05）, however, the root extract did not cause a significant difference in interleukin-10 levels compared to the control （P〉0.05）. CONCLUSION： The findings suggest that L involving protective effects against oxidative ported might be a potential immune-modulating agent damage.

Genes Expression Profile Difference in Peripheral Blood Between Esophageal Carcinoma Patients and Normal Subjects

Objective： To study the genes expression profile differences in the peripheral blood between esophageal carcinoma patients and normal subjects using the gene chip technique and screen out the esophageal early conceration associated genes. Methods： The total RNA was extracted and purified in the peripheral blood obtained from the patients with esophageal carcinoma and normal subjects. The first strand of cDNA was synthesized through retro-transcription and labeled with Cy5 and Cy3 fluorescence as probes. The mixed probes were hybridized with a piece of 4096 double dot human whole gene chip. The acquired image was analyzed by microarrav suite software using a digital computer, and the intensity of ttuorescence signal and its ratio were calculated. Results： A total of 92 genes were screened out and its expression difference was more than 2 times in the peripheral blood between the patients with esophageal carcinoma and normal subjects. Among these, the expression difference of 36 genes was more than 3 times. Two human urokinase plasminogen activator surface receptor （UPAR） genes, 80K-L protein gene, human protein tyrosine-phosphatase gent arid proto-oncogene protein mRNA were significantly up-regulated, while the collagen V type （α-2 gene was markedly down-regulated. Conclusion： 80K-L protein gene, tyrosinephophatase gene, proto-oncogene protein arid the collagen V type α-2 gene might be associated with the ontogenesis, development and its metastasis in the esophageal carcinoma. The UPAR gene may play important roles in the diagnosing the micrometastasis in the peripheral blood of esophageal carcinoma.